垂盆草饮片及其提取物高效液相色谱指纹图谱与多成分含量测定

HPLC Fingerprints and Multi-Component Content Determination of Decoction Slice and Extract of Sedi Herba

目的评价垂盆草饮片及其提取物的质量。方法采用高效液相色谱(HPLC)法测定垂盆草饮片中没食子酸、金丝桃苷、木犀草苷、槲皮素、山柰酚、异鼠李素的含量,色谱柱为Hanbon Hedera ODS-2 C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,检测波长为280 nm(没食子酸)和350 nm(金丝桃苷、木犀草苷、槲皮素、山柰酚、异鼠李素),柱温为35℃,进样量为20μL。建立15批垂盆草饮片及其提取物的HPLC指纹图谱,采用中药色谱指纹图谱相似度评价系统(2004A版)进行相似度评价,结合主成分分析法对垂盆草饮片及其提取物进行质量评价。结果15批垂盆草饮片中6种成分平均含量为0.0054%~0.0488%,RSD为3.05%~24.84%;15批垂盆草提取物中6种成分的平均含量为0.0323%~0.2725%,RSD为44.06%~77.20%。共确定20个共有峰,指认出峰1为没食子酸、峰9为金丝桃苷、峰10为木犀草苷、峰15为槲皮素、峰18为山柰酚、峰20为异鼠李素。垂盆草饮片相似度为0.739~0.942,垂盆草提取物相似度为0.735~0.998。主成分分析结果显示,垂盆草饮片及其提取物分别得到2个和3个主成分,累计方差贡献率分别为97.130%,94.648%;垂盆草饮片及其提取物主成分综合得分排序按样品编号从大至小分别为S2>S1>S3>S4>S5>S6>S9>S7>S8>S10>S12>S13>S11>S14>S15,T14>T12>T15>T13>T11>T7>T9>T1>T2>T8>T3>T10>T4>T6>T5。结论所建立的方法简便快速、稳定可靠,可用于垂盆草饮片及其提取物的质量控制与评价。

Objective To evaluate the quality of decoction slice and extract of Sedi Herba.Methods High-performance liquid chromatography (HPLC) method was used to determine the contents of gallic acid,hypericin,luteolin,quercetin,kaempferol,and isorhamnetin in the decoction slice and extract of Sedi Herba.The chromatographic column was Hanbon Hedera ODS-2 C_(18)column (250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.1%phosphoric acid aqueous solution (gradient elution),the flow rate was 1.0 mL/min,the detection wavelengths were 280 nm (gallic acid) and 350 nm (hypericin,luteolin,quercetin,kaempferol,and isorhamnetin),the column temperature was 35℃,and the injection volume was 20μL.The HPLC fingerprints of 15 batches of Chinese herbal medicine decoction pieces and their extracts were established,and the similarity evaluation was conducted by the Similarity Evaluation System for Fingerprint Chromatographic of Traditional Chinese Medicine(Version 2004).The quality of decoction slice and extract of Sedi Herba was evaluated by the principal component analysis(PCA).Results The average content of six components in 15 batches of decoction slice of Sedi Herba was in the range of 0.0054%-0.048 8%,with RSD of 3.05%-24.84%.The average content of six components in 15 batches of extract of Sedi Herba was in the range of 0.032 3%-0.272 5%,with RSD of 44.06%-77.20%.A total of 20 common peaks were identified,with peak 1 identified as gallic acid,peak 9 as hypericin,peak 10 as luteolin,peak 15 as quercetin,peak 18 as kaempferol,and peak20 as isorhamnetin.The results of PCA showed that there were two principal components in the decoction slice of Sedi Herba and three principal components in the extract of Sedi Herba,with cumulative variance contribution rates of 97.130%and 94.648%,respectively.The comprehensive scores of the principal components in the decoction slice and extract of Sedi Herba were ranked in descending order of sample number as S2>S1>S3>S4>S5>S6>S9>S7>S8>S10>S12>S13>S11>S14>S15,T14>T12>T15>T13>T11>T7>T9>T1>T2>T8>T3>T10>T4>T6>T5.Conclusion The established method is simple,rapid,stable,and reliable,which can be used for quality control and evaluation of the decoction slice and extract of Sedi Herba.